AUTHOR=Hunziker Johan , Nishida Keiji , Kondo Akihiko , Ariizumi Tohru , Ezura Hiroshi 

TITLE=Phenotypic Characterization of High Carotenoid Tomato Mutants Generated by the Target-AID Base-Editing Technology

JOURNAL=Frontiers in Plant Science

VOLUME=13

YEAR=2022

URL=https://www.frontiersin.org/journals/plant-science/articles/10.3389/fpls.2022.848560

DOI=10.3389/fpls.2022.848560

ISSN=1664-462X

ABSTRACT=<p>Our previous study demonstrated that Target-AID which is the modified CRISPR/Cas9 system enabling base-editing is an efficient tool for targeting multiple genes. Three genes, <italic>SlDDB1</italic>, <italic>SlDET1,</italic> and <italic>SlCYC-B,</italic> responsible for carotenoid accumulation were targeted, and allelic variations were previously obtained by Target-AID. In this research, we characterized the effect of new alleles on plant growth and fruit development, as well as carotenoid accumulation, individually in segregating backcross populations or combined in null self-segregant lines. Only lines carrying homozygous substitutions in the three targeted genes and the segregating backcross population of individual mutations were characterized, resulting in the isolation of two allelic versions for <italic>SlDDB1,</italic> one associated with <italic>SlDET1</italic> and the last one with <italic>SlCYC-B.</italic> All edited lines showed variations in carotenoid accumulation, with an additive effect for each single mutation. These results suggest that Target-AID base-editing technology is an effective tool for creating new allelic variations in target genes to improve carotenoid accumulation in tomato.</p>